Kim, Se-Woong;Jung, Yeon-Gil;Park, Jong-Im;Roh, Sangho
Journal of Embryo Transfer
/
v.29
no.2
/
pp.127-132
/
2014
This study was carried out to investigate the effects of tissue inhibitor of matalloproteinase-1 (TIMP-1), Activin A and Heparin binding epidermal growth factor (HB-EGF) on in vitro production of bovine embryos. In experiment 1, presumptive zygotes were cultured in the medium supplemented with TIMP-1 ($0.5{\mu}g/ml$), Activin A (100 ng/ml), or HB-EGF (100 ng/ml) at $39^{\circ}C$ in a humidified atmosphere of 5% (v/v) $CO_2$, 5% (v/v) $O_2$ and 90% (v/v) $N_2$. In experiment 2, TIMP-1 + HB-EGF or Activin A + HB-EGF combinations were supplemented in the culture medium. The developmental rate to blastocysts, hatching rate and total cell numbers of the blastocysts were evaluated in both experiments. The embryos cultured in medium without growth factor supplementation was used as control group. In experiment 1, the embryos cultured in medium supplemented with TIMP-1 and Activin A showed significantly higher developmental rate to blastocysts than those cultured with HB-EGF and control (36.9%, 34.1%, 21.2% and 23.1%, respectively) (P<0.0001). However, the hatching rate of blastocyst was significantly higher in embryos with HB-EGF than those with TIMP-1, Actvin A and Control groups (84.4%, 58.8%, 51.4% and 49.3%, respectively) (P<0.001). Total cell number per blastocyst was also significantly higher in embryos with HB-EGF group ($174.3{\pm}2.5$) than those with TIMP-1, Activin A (149.7 and 150.0, respectively) (P<0.05) and Control (119.0) (P<0.001). In experiment 2, embryos cultured with combined treatment of Activin A and HB-EGF resulted in significantly higher rates of blastocysts formation (48.0%), hatching rate (89.7%) and total cell number in blastocyst ($182.3{\pm}2.1$) than those with TIMP-1 and HB-EGF combination group (32.0%, P<0.001; 76.6%, P<0.05; $165.7{\pm}4.2$, P<0.001, respectively). Our data demonstrate that in vitro production of bovine embryos could be improved by combined supplementation of Activin A and HB-EGF in culture medium.
Nasal-type extranodal natural killer (NK)/T-cell lymphoma (ENKL) is a highly invasive cancer with a poor prognosis. More effective and safer treatment regimens for ENKL are needed. Pegaspargase (PEG-Asp) has a similar mechanism of action to L-asparaginase (L-Asp), but presents lower antigenicity. The aim of the present research was to evaluate the safety profile and the latent efficacy of a PEG-Asp-based treatment regimen in patients with ENKL. Data collected from 20 patients with histologically confirmed ENKL, admitted to the Third Affiliated Hospital of Sun Yat-Sen University from January 2009 to August 2013, were included in the study. All patients received $2500IU/m^2$/IM PEG-Asp on day 1 of every 21-day treatment cycle. Patients received combination chemotherapy with CHOP (n=5), EPOCH (n=7), GEMOX (n=7) or CHOP with bleomycin (n=1). After 2-5 treatment cycles (median, 4 cycles) of PEG-Asp-based chemotherapy, five patients (25%) showed a complete response (CR), and the overall response rate (ORR) was 60%. Grade 3/4 neutropenia occurred in fourteen patients (70%). Grade 3 alanine aminotransferase (ALT) elevation was observed in two. Grade 1-2 non-hematological toxicity consisted of activated partial thromboplastin time (APTT) elongation (n=9), hypofibrinogenemia (n=6), hypoproteinemia (n=17), hyperglycemia (n=3), and nausea (n=6). No allergic reactions were detected. No treatment related death was reported. Our results suggested that PEG-Asp-based chemotherapy presented an acceptable tolerance and a potential short-term outcome in patients with nasal-type ENKL.
Park Gyeong-Seon;Jang Yeon-Jin;Park Chun-Sik;Im Chae-Heon
Proceedings of the Korean Biophysical Society Conference
/
1999.06a
/
pp.61-62
/
1999
;The mechanisms inducing hypertension are actively investigated and are still challenging topics. Basically hypertension must be caused by the disorder of $Ca^{2+}$ metabolism in vascular smooth muscle, such as the increase of $Ca^{2+}$ influx, the decrease of ci+ efflux, or the change of sensitivity of contractile protein etc. The one of cause of the increase of ci+ influx may be the change of ci+ channel activity. Even though the relationships of ci+ channel activity and hypertension were studied using various hypertension models, still it is not clear how much change of $Ca^{2+}$ channel activity in diabetes mellitus (DM) induced hypertension is occurred. We induced DM hypertension in SD rat and compared the $Ca^{2+}$ channel activity with age-matched normotensive SD rat. For inducing DM hypertension, left kidney was removed with 200 gm rat and, after 1 month, 60 mg/kg of streptozotocin was injected into peritoneal space to induce diabetes mellitus. Usually after 4-6 weeks, hypertension was fully induced. For isolating vascular smooth muscle cells (VSMC), we used mesenteric arteriole (3rd - 4th branch of mesenteric artery) of which diameter is below 150 urn. VSMCs were isolated enzymatically. $Ca^{2+}$ current was measured using whole cell patch clamp technique. All experiments were performed at $37^{\circ}C$. The cell membrane area of VSMC of DM hypertensive rat is larger than that of control VSMC($36.6{\pm}3.64{\;}pF{\;}vs{\;}22.4{\pm}1.29{\;}pF, {\;}mean{\pm}S.E.$) When we compared the current amplitude, the $Ca^{2+}$ current amplitude in VSMC of DM hypertensive rat is much larger than that in VSMC of normotensive age-matched rat. After $Ca^{2+}$ current amplitude was normalized by cell membrane area, the current amplitude in DM hypertension is increased to $249.1{\pm}15.9{\;}%{\;}(mean{\pm}S.E.M)$, which means the ;absolute current amplitude is about 4 times larger in DM hypertension. When we compared the steady state activation and inactivation. there were no noticeable differences. From these results. one of cause of the DM hypertension is due to the increase of $Ca^{2+}$ current amplitude. But it need further study why the $Ca^{2+}$ current is so large in VSMC of DM hypertension and how much $Ca^{2+}$ influx through $Ca^{2+}$ channel contribute to the increase of intracellular $Ca^{2+}$ and eventually contribute to development of hypertension.ypertension.
Jung Eun Kim;Bo-Ram Kim;Su Hui Seong;Jin-Ho Kim;Ha-Nul Lee;Chan Seo;Ji Min Jung;Su A Im;Kyung-Min Choi;Jin-Woo Jeong
Proceedings of the Plant Resources Society of Korea Conference
/
2023.04a
/
pp.50-50
/
2023
Prostaglandin E2(PGE2), a major product of cyclooxygenase-2 (COX-2), plays an important role in the carcinogenesis of many solid tumors, including colorectal cancer. Because PGE2 functions by signaling through PGE2 receptors (Eps), which regulate tumor cell growth, invasion, and migration, there has been a growing amount of interest in the therapeutic potential of targeting Eps. In the present study, we investigated the role of EP4 on the effectiveness of cordycepin in inhibititing the migration and invasion of HCT116 human colorectal carcinoma cells. Our data indicate that cordycepin suppressed lipopolysaccharide (LPS)-enhanced cell migration and invasion through the inactivation of matrix metalloproteinases (MMP)-9 as well as the down-regulation of COX-2 expression and PGE2 production. These events were shown to be associated with the inactivation of EP4 and activation of AMP-activated protein kinase (AMPK). Moreover, the AMPK inhibitor, compound C, as well as AMPK knockdown via siRNA, attenuated the cordycepin-induced inhibition of EP4 expression. Cordycepin treatment also reduced the activation of CREB. These findings indicate that cordycepin suppresses the migration and invasion of HCT116 cells. Through modulating EP4 expression and the AMPK-CREB signaling pathway. Therefore, cordycepin has the potential to serve as a potent anti-cancer agent in therapeutic strategies against colorectal cancer metastasis.
The study was carried out to evaluate the anticancer effects in Korean soybean paste(doenjang) added with sun-dried salt(S-D), roasted salt(R-D), bamboo salt roasted once(B1-D) or bamboo salt roasted nine times(B9-D). In MTT assay, S-D, R-D, B1-D and B9-D exhibited a significant inhibitory activity(64-87% and 68-92%) against the proliferation of AGS human gastric adenocarcinoma cells by adding 0.05% and 0.1% of methanol extract. Among Balb/c mice injected with sarcoma-180 cells, the bodies, livers, spleens, kidneys and heart weight of the mice administered with 4 kinds of doenjang extracts were heavier than the non-administered ones of the group. However, no difference was found between the control and doenjang administered groups. Four kinds of doenjang inhibited significantly the tumor growth, especially R-D and B1-D showing an inhibition of tumor cell growth up to 97% by the administration of 1.0 mg/kg methanol extracts of doenjang. The activity of natural killer(NK) cells was relatively high in mice administered with four kinds of doenjang. Particularly, mice administered with R-D methanol extract showed a stronger activity of 88.2%. The activity of glutathione S-transferase(GST) in mice administered with four kinds of doenjang was higher than that of the non-administered group. In particular, the GST activity was the strongest in the group with B1-D. As these results indicate the various kinds of salt have different effects on the anticancer activity of doenjang.
Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
Journal of Mushroom
/
v.20
no.3
/
pp.178-182
/
2022
Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.
Background : Various combinations of treatment modalities have been reported in stage III non-small cell lung cancer (NSCLC). however, the standard treatment modality has not established yet. Recently, the efficacy of concurrent chemotherapy and radiation therapy has been reported in locally advanced lung cancer. We evaluate the response rate, toxicity, and survival of concurrent chemotherapy with etoposide and cisplatin(EP) and radiation therapy for unresectable stage III NSCLC. Method : Between October 1995 and December 1996, 32 patients with histologically proven unresectable stage III NSCLC without malignant pleural effusion were entered into this study. Twenty-nine patients were eligible for the response, survival, and toxicity analysis. Induction was two cycles of chemotherapy with etoposide and cisplatin plus concurrent chest RT to 4500cGy. Resection was attempted if the clinical response offered surgical resectability. Boost radiation therapy upto 5940cGy and one cycle of EP were performed if the disease were stable or responsive but still unresectable. Results : Of 29 eligible patients, 22(75.9%) showed partial response(PR). The progression free interval was 6.3months(range 1.1 to 19.5months). Surgical resection was performed in one patient. The median survival was 12.1months and one-year survival rate was 50.6%. The major toxicity was leukopenia($\geq$ grade 3, 46%). Thrombocytopenia over grade 3 was found in 11%. Radiation pneumonitis occurred in 13 patients(46%). Conclusion : Concurrent chemotherapy(EP) plus radiotherapy was effective and tolerable in the treatment of unresectable stage III NSCLC.
The optimum dosage of quicklime in producing organic fertilizer using livestock wastes vith a greater than 80% water content was analysed. After one day had elapsed to allow for the organic fentilizer to dry, the quicklime dosage and the composition of the organic fertilizer were analysed. Any from done to the organic fertilizer was also assessed. The amount of the quicklime required to stabilize livestock wastes was determined by water content of livestock wastes. For J farm(slurry style) of which livestock wastes have 94.6% of water concentration, less than 3% of total amount of livestock wastes, for H farm (scraper style) of which livestock wastes have 85% of water concentration, less then 4% of total livestock wastes and Y farm(traditional style) of which livestock wastes have 80% of water concentration, less then 5% of total livestock wastes. Generally, in order to pack the organic fertilizer, water containing quicklime0stabilized livestock wastes should be less than 35%. It takes 9 days to keep this water content for the wastes from H and Y farms(less than 85% in water content), and 12 days for the wastes from J (94.6% in water content). According to the classification standard for compost constitution by Higgins, the crude fertilizers from all 3 farms had high grade $K_2O$ and CaO, the middle grade T-N and middle or low grade $P_2O_5$. Stabilization by quicklime is known to inhibit bacterial decomposition of organic matter and the activity of pathogenic organisms. In this study, more then 99.99% of coliform group, fecal group and viable cell count were reduced. Our results indicate that livestock wastes of greater 80% water content could be used to produce organic fertilizer without the addition of a material for moisture control.
The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.
Youn, Seok Jea;Kim, Hun Nyun;Im, Jong Kwon;Kim, Yong-Jin;Baek, Jun-Soo;Lee, Su-Woong;Lee, Eun Jeong;Yu, Soon Ju
Journal of Environmental Science International
/
v.26
no.8
/
pp.913-925
/
2017
Phytoplankton populations were examined at three sites in Lake Cheongpyeong, South Korea from March 2008 to December 2016, including measurement of phytoplankton communities and their dominant species, abundance and environmental factors. The annual average ranges of water temperature, dissolved oxygen and conductivity were $15.2-18.8^{\circ}C$, 10.3-12.2 mg/L, $86-140{\mu}S/cm$, respectively, with similar values at all studied sites. The highest phytoplankton cell density was observed in spring and fall, and it subsequently decreased rapidly during heavy rainfall. Diatoms were dominant in spring (mainly Stephanodiscus hantzschii, Asterionella formosa) and fall (mainly Aulacoseira granulata), while greenalgae and cyanobacteria had high appearance in early-summer and summer, respectively, indicating that water temperature is the most important factor influencing their growth. Stephanodiscus hantzschii and Asterionella formosa frequently occurred at low water temperature ($4.5-15.0^{\circ}C$ and $5.4-21.6^{\circ}C$, respectively) while Aulacoseira granulata and Anabaena spp. were favored by high water temperature (8.6-28.4 and $14.9-26.2^{\circ}C$, respectively) and phosphorus. Additionally, Fragilaria crotonensis occurred at low nutrient conditions. Rhodomonas spp. frequently appeared year-round.
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